龙须菜四分孢子体cDNA文库的构建  

Construction of a cDNA Library from the Tetrasporophyte ofGracilar ia lemaneiformis (Rhodophyceae)

作　　者：孙雪[1] 张学成[1] 隋正红[1] 茅云翔[1] 

机构地区：[1]中国海洋大学海洋生命学院,青岛266003

出　　处：《青岛海洋大学学报（自然科学版）》2003年第5期727-732,共6页Journal of Ocean University of Qingdao

基　　金：国家自然科学基金项目 ( 30 170 736 )资助

摘　　要：为了研究不同世代和性别基因表达谱的差异 ,本文构建了龙须菜 (Gracilarialemaneiformis)四分孢子体的 c DNA文库。总 RNA提取使用 RNeasy Plant Mini Kit(Qiagen) ,c DNA文库构建使用 SMART c DNA Library Construction Kit(Clontech) ,包装蛋白购自 Promega公司。该c DNA文库含有 1 .2 8× 1 0 6个克隆 ,扩增文库的滴度是 1 .98× 1 0 9pfu/ m L,重组频率是 85.0 % ,插入片段几乎全部集中在 50 0～ 1 50 0In order to study th e differentially expressed genes in different phases and sexes of Gracilaria l emaneiformis, a cDNA library from the tetrasporophytic thalli was constructed. Total RNA was isolated by using RNeasy Plant Mini Kit from Qiagen. The cDNA lib rary was constructed by SMART cDNA Library Construction Kit from Clontech, while the packing system was purchased from Promega. The constructed cDNA library con tains 1.28×10 6 independent clones, the titer of the amplified library is 1.98 ×10 9 pfu/mL and the recombinant rate is 85.0% calculated by the amplification of randomly selected 20 clones. Of the amplified 17 clones, 16 insert DNAs rang e from 500 bp to 1500 bp in length and one is about 450 bp.

关 键 词：龙须菜 四分孢子体 CDNA文库 基因表达谱 分子生物学 

分 类 号：Q943.2[生物学—植物学] Q344.13